J. Org. C h e m . 1987,52, 4792-4796
4792
Acknowledgment. This research was supported by the Comisidn Asesora de Tnvestigacidn Cientifica y TBcnica (CAICYT Project 904/84). Registry No. 1,3068-88-0;2,625-38-7; 3, 107-93-7;5,96-48-0;
Scheme In
6, 3724-55-8;7, 623-43-8.
Supplementary Material Available: Reaction rates for acid-promoted transformations under heating are summarized in Figures 1-3 (4 pages). Ordering information is given on any current masthead page.
Studies on the Syntheses of Sesquiterpene Lactones. 10.' Improved Syntheses of ( )-Tuberiferin and the Related &-Methylene y-Lactones and Their Biological Activities
+
Masayoshi Ando,*% Tetsuo Wada,28 Haruhiko Kusaka,2* Kahei Takase? Naonori Hirata,2b and Yoshikazu Yanagi2c Department of Chemistry, Faculty of Science, Tohoku University, Aramaki-aza-Aoba, Sendai 980, Japan, Pesticides Research Laboratory, Sumitomo Chemical Company, L t d . , and Research Laboratories, Sumitomo Pharmaceutical Company, L t d . , Takatsuka, Takarazuka 665, J a p a n
0
87% overall yield from (7 without purification of 8)
53.5% overall
Received January 15, 1987
Sesquiterpene lactones with an a-methylene y-lactone moiety fused on various skeletons are a rapidly expanding group of natural products, comprising to date more than 900 ~ a r i e t i e s . ~Some of them have been shown to have considerable biological activities as allergenic agents, cytotoxic and antitumor agents, regulators of plant growth and antimitotic activity, and antishistosomal agents. Because of their biological activities and because they are available from natural sources often only in small quantities, their efficient syntheses are a synthetic challenge that has received much attention and many reports of such syntheses have appeared in the last decade. In the course of our program of the study of the structure-activity relationship of a-methylene y-lactones, we needed efficient syntheses of (+)-tuberiferin (9) and related a-methylene y-lactones in gram quantities. Although total syntheses of (&)- and (+)-tuberiferin (9) had already appeared in the l i t e r a t ~ r ethe , ~ reported methods were inconvenient for our purpose. In the reported total syntheses of (&)- and (+)-9 the introduction of the a-methylene y-lactone moiety was left to the final stage, probably because of its expected reactivity and instability. Since we had noticed that the a-methylene y-lactone moiety was stable under acidic and mild basic conditions and bromination by phenyltrimethylammonium perbromide (PTAB), we envisioned another approach to 9 in which, as shown in Scheme I, the introduction of the amethylene y-lactone moiety was at an early stage. In this note we report efficient syntheses of (+)-tuberiferin (9) and the related a-methylene y-lactones 6-8 and their biological activities. Part 9 Ando, M.; Ono, A.; Takase, K. Chem. Lett. 1984, 493. (a) Department of Chemistry, Faculty of Science, Tohoku University. (b) Pesticides b e a r c h Laboratory, Sumitomo Chemical Co., Ltd. (c) Research Laboratories, Sumitomo Pharmaceuticals Co., LM. (3) Fisher, N. H.; Olivier, E. J.; Fisher, H. D. Prog. Chem. Org. Nut. (1) (2)
Prod. 1979, 38, 214.
(4) Two syntheses of this compound have already been reported; (a) Yamakawa, K.; Nishitani, K.; Tominaga, T. Tetrahedron Lett. 1976,2829. (b) Grieco, P. A.; Nishizawa, M. J . Chem. SOC.,Chem. Commun. 1976, 582.
0022-3263/87/1952-4792$01.50/0
(+I-Tuberiferin
(
9
I
)
eudesnane
(10 1
Reagents and conditions: (a) H,/2% Pd-SrC03, AcOEt; (b) HC1, EtOH; (c) HO(CH2),0H, p-TsOH, benzene, reflux; (d) LDA, THF; (e) (C8H6Se),,HMPA, THF; (f) 30% H,Oz, AcOH, THF; (9) 20% aqueous AcOH-EtOH, reflux; (h) 1.1 equiv of PTAB, THF; (i) Li,C03, LiBr, DMF, 123-131 OC; (i) 50% aqueous AcOH, reflux. (I
Results and Discussion Synthesis of (+)-Tuberiferin (9). The starting material 3 was obtained from commercially available l-asantonin (1) by modification of the known procedure5 (Scheme I). Thus, catalytic hydrogenation of 1 over 2% palladium on strontium carbonate in ethyl acetate and epimerization of the resulting 2 possessing a p(axia1)methyl group at C4 by 2 M hydrochloric acid in ethanol gave 3 in 81% yield. Acetalization of the C,-carbonyl group of 3 under standard conditions gave acetal 4 in 85% yield. Phenylselenenylation of 4 by Grieco's method6 gave the corresponding phenyl selenide ( 5 ) in 96% yield. Treatment of 5 with 30% hydrogen peroxide in THF in the presence of acetic acid gave an a-methylene y-lactone (6) in 93% yield. Deacetalization of 6 was achieved by treatment with 20% aqueous acetic acid in ethanol at 85 OC for 4 h to give 7 in a quantitative yield. 7 was also prepared by a different procedure. Thus, deacetalization of 5 in boiling 50% aqueous acetic acid gave the corresponding keto selenide 10 in 97% yield. Successive treatment of 10 with 30% hydrogen peroxide in THF in the presence of acetic acid gave 7 in 93% yield. The selective bromination at C2 of 7 was achieved as follows. Treatment of 7 with 1.05 mol equiv of PTAB' in (5) (6)
Corey, E. J.; Hortmann, A. G.; J . Am. Chem. SOC.1965,87,5736. Grieco, P. A.; Miyashita, M. J. Org. Chem. 1974, 39, 120.
0 1987 American Chemical Society
J. Org. Chem., Vol. 52, No. 21, 1987 4793
Notes Table I. Cell Growth Inhibitory Activity against Murine Lymphocytic Leukemia (P388) in Vitro cell growth inhibitory ratio: %
Table 11. Plant Growth Inhibitory Activities of Compounds 6-9'
compd 6 4
7 3 8 11 9 12 adriamycin (control)
105 22 102 16 103 103 107 76 107
98 5 40 7 106 69 102 14 103
6 16 0 5 80 13 33 1 104
plant E . frumentacea
B. juncea C . sativus E . frumentacea
B. juncea 35
C . sativus
"Cell growth inhibitory ratio (%) = [l - (T- C,)/(C - C,)] X 100 where T = cell cound after culture with compound, C = cell count after culture without compound, and C, = cell count a t the start of culture.
B. juncea
Chart I
C. sativus
E. frumentacea
E. frumentacea B. juncea C. sativus
THF at -6 OC for 1h gave an a-bromo ketone (8)in 86% yield. The bromine atom at C2 of 8 was a-equatorial because of the magnitude of the coupling constant of the C2 proton [6 4.83 (1H, ddd, J = 12.8, 6.5, 1.2 Hz)].* Dehydrobromination of 8 with lithium carbonate and lithium bromide in DMF at 123-131 "C for 100 min gave am a,@-unsaturatedketone (9) in 85% yield. 9 was also obtained in 87% overall yield from 7 by using the crude bromination product without any further purification. 9 was identical with (+)-tuberifering by direct comparison of their 'H NMR spectra (60 MHz, CDCI3).I0 The IR spectrum, melting point, and [a]Dvalue of 9 were also in good accordance with those of (+)-tuberiferin reported in the literaturea4sg Biological Activities. 1. Cell Growth Inhibitory Activity toward the P388 Lymphocytic Leukemia Test System. The a-methylene y-lactones 6-9 showed significant cell growth inhibitory activity against murine lymphocytic leukemia (P388) in vitro. The results are compared with those of the corresponding a-methyl ylactones in Table I. As expected, the a-methylene ylactones 6-9 showed stronger activity than the corresponding saturated a-methyl y-lactones 4, 3, 11, and 12 (Chart I). It is interesting that the a-bromo ketone (11) possessing a saturated a-methyl y-lactone moiety showed significant cell growth inhibitory activity against P388 lymphocytic leukemia in vitro. 2. Plant Growth Regulating Activity. The plant growth regulating activity of a-methylene y-lactones 6-9 was studied employing three kinds of seeds, Echinochloa frumentacea (Japanese millet, Japanese name shokuyb hie), Brassica juncea (brown mustard, Japanese name (7)Fieser, M.; Fieser, L. F. Reagent for Organic Synthesis; Wiley: New York, 1975;Vol. 5, pp 531-532. (8)(a) Bhacca, N. S.; Williams, D. H. Application of NMR Spectroscopy in Organic Chemistry; Holden-Day: San Francisco, 1964;pp 73-77. (b) Jackman, L.M.; Sternhell, S. Application of Nuclear Magnetic Resdnance Spectroscopy in Organic Chemistry, 2nd ed.; Pergamon: Oxford, 1969;pp 334-341. (9)Barrera, J. B.; BretBn, T. L.; Fajardo, M.; GonzHlez, A. G. Tetrahedron Lett. 1967,3475. (10)Yoshioka, H.; Mabry, T. J.; Timmermann, B. N. Sesquiterpene Lactones; University of Tokyo: Tokyo, 1973;p 236.
part stem root stem root stem root stem root stem root stem root stem root stem root stem root stem root stem root stem root
activities 1OOOppm 100ppm
+
++ - to + +-
+ +++ ++
+ to ++
++ to +++ + +++ ++ + to- ++ + - to
+++ +++ ++ to +++ +++ ++ to +++ +++
OKey: +++, serious or complete inhibition of seeds germination and seedling growth; ++, obvious effect on seeds germination and seedling growth; +, slight detectable effect; -, no effect.
seiyb karashina), and Cucumis sativus (cucumber, Japanese name kyari). The germination of seeds and growth of seedlings were observed. Compounds 7 and 9 showed significant plant growth inhibitory activity at 1000 ppm. The results are summarized in Table 11. 3. Control of Crop Diseases. The preventive and curative activities of compounds 4 and 6-9 in controlling crop diseases were examined by pot test. The a-methylene y-lactones 7-9 showed significant preventive activities in controlling dawny mildew of grape and late blight of tomato. The results are summarized in Table 111;
Experimental ,Section All melting points were uncorrected. lH and 13CNMR spectra were recorded in CDC13. Mass spectra were recorded a t 25 eV. (llS)-3-Oxoeudesmano-13,6a-lactone (3). A mixture of l-a-santonin (1; 23.10g, 93.79 mmol), ethyl acetate (750 mL), and 2% palladium on strontium carbonate (10.3g) was shaken under 1 atm hydrogen at room temperature. The hydrogen uptake ceased at 5.1 L (227.68mmol) in 41 min. The catalyst was removed, and the filtrate was concentrated to 100 mL to give the first crop of (11S)-3-oxo-4&-eudesmano-13,6a-lactone(2,10.95 9). The filtrate was further concentrated to give the second crop of 2 (5.28g), which was indistinguishable from the first crop in the comparison of their NMR and IR spectra. The combined yield of 2 was 69.1%. The mother liquid was concentrated to give an oily crude material (10.97 g), which was dissolved in a mixture of ethanol (60 mL) and 2 M HCl (6 mL) and allowed to stand at room temperature for 15 h. The mixture was poured into a saturated aqueous solution of NaCl(l50 mL) and extracted with ethyl acetate (4 X 40 mL). The combined extracts were washed with a saturated aqueous solution of NaCl, dried (Na2S04),and concentrated to give a crude crystalline material (3.56 g), which was recrystallized from ethanol to give 3 (2.71g, 11.5%) as needles. The combined 2 (16.23g) obtained above was allowed to stand at room temperature in a mixture of ethanol (60mL) and 2 M HC1 (9 mL) for 15 h and worked up in the above mentioned manner to give 3 in a quantitative yield. Thus, the combined yield of 3 from 1 was 81%. The analytical sample of 2 was obtained by recrystallization from ethyl acetate as plates: mp 141.5-144.5 OC (lit.5 mp 145.5147 "C); IR (KBr) 1765,1708cm-'; 'H NMR (60 MHz) 6 1.20 (3 H,
4794
J. Org. Chem., Vol. 52, No. 21, 1987
comDd 4 6
7 8 9
Notes
Table 111. Preventive or Curative Activities of Compounds 4 and 6-9 in Controlling Crop Diseases evaluation of disease controla concn, blast of sheath blight powdery mildew damping off downy mildew late blight DDm riceb of ricec of wheatd of cucumbe? of grape' of tomato8 0 0 0' 500 1 0 0 0' 500 3 4 0 0' 500 2 4 0 200 1 0 50 4 3 0' 500 2 4 4 200 0 0 50 4 0 0' 500 1 2 4 200 1 0 50
scap of appleh 0 0 0 0
1
"This assessment was made by disease severity in rice, wheat, grape, tomato, and apple or numbering infecting seedlings of cucumber. The indices are expressed on a scale of five: 5, 100%; 4,99-90%, 3, 89-70%, 2, 69-50%, 1, 49-30%, 0, 2 9 4 % . bCaused by Pyricularia oryzae. Caused by Rhizoctonia solani. Caused by Erysiphe graminis. e Caused by Pythium aphanidermatum. f Caused by Plasmopara uiticora. #Caused by Phytophthora infestano. Caused by Ventria inaequalis. Curative activity. The preventive activity was not determined at this concentratibn.
'
CiO-CH3), 1.23 (3 H, d, J = 6.5 Hz), 1.24 (3 H, d, J = 7.5 Hz), 4.00 (1H, dd, J = 10.0, 10.0 Hz, CG-H).Anal. Calcd for C&&$ C, 71.97; H, 8.86. Found: C, 71.99; H, 8.61. The analytical sample of 3 was obtained by recrystallization from a mixture of chloroform and ether (1:l)as needles: mp 153-155 "C (lit." mp 154-155 "C); IR (KBr) 1775,1698 cm-'; 'H NMR (60 MHz) 6 1.19 (3 H, s, Clo-CH3),1.22 (3 H, d, J = 7.2 Hz), 1.25 (3 H, d, J = 7.0 Hz), 3.91 (1H, dd, J = 11.0,g.O Hz, C&). Anal. Calcd for C15H2203: C, 71.97; H, 8.86. Found: C, 71.79; H, 9.06. (115)-3,3-(Ethylenedioxy)eudesmano-13,6a-lactone (4). A mixture of 3 (1.00 g, 3.99 mmol), ethylene glycol (5 mL, 89.4 mmol), and p-toluenesulfonic acid (40.2 mg, 0.21 mmol) in dry benzene was refluxed in a flask equipped with a Dean-Stark column packed with molecular sieves for 24 h. The mixture was cooled, diluted with a saturated aqueous solution of NaCl(100 mL), and shaken, and the benzene layer was drawn off. The aqueous layer was further extracted with benzene (4 X 10 mL). The combined extracts were washed with a saturated aqueous solution of NaCl (3 X 10 mL), dried, and concentrated to give a crystalline crude product, which was recrystallized from a mixture of benzene, ether, and hexane (1:l:l)to give 4 (998 mg, 85%) as colorless prisms: mp 167-168 "C; IR (KBr) 1770 cm-'; 'H NMR (60 MHz) 6 0.99 (3 H, 9, Cio-CH,), 1.04 (3 H, d, J = 6.2 He, Cd-CHS), 1.18 (3 H, d, J = 6.2 Hz, Cii-CHB), 3.81 (1H, dd, J = 10.5, 10.5 Hz, C,-H), 3.93 (4 H, br s, OCH2CH20);13C NMR 6 12.0 (9, C15), 12.5 (q, c12), 18.1 (9, ci4),23.1 (t, c8),31.1 (t, c2),36.6 (9, clo),39.0 (t, Cg), 39.4 (d, C4),40.6 (d, Cl1), 40.6 (t,Cl), 50.9 (d, C5 or Cd, 53.5 (d, C5 or C7),65.1 (t, OCH2CH20),83.2 (d, c6),110.5 ( 8 , c3), 179.3 (s, C13); MS, m/e (relative intensity) 294 (M', 6.9), 99 (100); ['YI2OD +24.3" (c 1.14, CHC13). Anal. Calcd for C17H2604: C, 69.36; H, 8.90. Found: C, 69.56; H, 9.04. S,
11~-(Phenylseleno)-3,3-(ethylenedioxy)eudesmano-13,6alactone (5). To a T H F solution of lithium diisopropylamide [prepared from diisopropylamine (0.28 mL, 2.00 mmol), 1.50 M butyllithium in hexane (1.48 mL, 2.22 mmol), and THF (5.0 mL) at -78 "C] was added dropwise over a period of 1h 296 mg (1.00 mmol) of 4 in T H F (7.5 mL). After the solution was stirred a t -78 "C for 75 min, diphenyl diselenide (625 mg, 2.00 mmol) in T H F (50 mL) containing HMPA (348 mg, 2.00 mmol) was added dropwise a t -78 "C over a period of 50 min. The reaction mixture was stirred a t -78 "C for 30 min and then warmed to -40 "C, where stirring was continued for an additional 45 min. The reaction was quenched by the addition of 0.1 M aqueous solution of HC1 (25 mL). The mixture was extracted with chloroform (3 X 30 mL). The combined extracts were washed with a saturated aqueous solution of NaCl (2 X 30 mL), dried, and concentrated to give a crude product, which was subsequently chromatographed over silica gel (Merck, 70-230 mesh, 50 g) and eluted with chloroform to give diphenyl diselenide. The fractions eluted with ethyl acetate gave spectroscopically pure 5 (432 mg, 96%) 89 pale yellow prisms. (11) Cocker, W.; McMurry, T. B. H. J. Chem. SOC. 1956, 4549.
The analytical sample was obtained by recrystallization from a mixture of benzene and ether (1:l)as pale yellow prisms: mp 180.5-181.5 O C ; IR (KBr) 1770 cm-'; 'H NMR (90 MHz) 6 1.01 (3 H, S, ClO-CH3), 1.04 (3 H, d, J = 6.7 Hz, Cd-CHJ, 1.50 (3 H, s, C11-CH3),2.01 (1H, dq, J = 11.3, 6.7 Hz, C4-H),3.93 (4 H, br S, OCH2CH20), 4.24 (1 H, dd, J = 9.8, 9.8 Hz, Ce-H), 7.18-7.38 (3 H), 7.52-7.65 (2 H); 13CNMR 6 12.0 (9, C15), 18.1 (9, Cu), 21.0 (t,C8), 22.3 (4, C12)t 31.1 (t, C2), 36.5 (8, Cia), 38.8 (t,GI),39.4 (4 C4), 40.5 (t, CJ, 48.7 (9, CiJ, 51.0 (d, Cb), 57.8 (d, C7), 65.1 (t, OCH2CH20), 81.1 (d, C6), 110.4 ( 5 , C3), 124.4 (S, CsH5), 128.9 (d, CsH5), 129.5 (d, C6H5),138.1 (d, C6H5),176.6 (8, c13); MS, m/e (relative intensity) 452 [M' (s2Se),11,450 [M' (@%e), 41,448 [M' (78Se),31,447 [M+ (77Se),11,446 [M' (76Se),11, 292 (77), 99 (100). [.IND +33.1° (c 2.00, CHC13). Anal. Calcd for C2,H3,O4Se: C, 61.46; H, 6.73. Found: C, 61.51; H, 6.90.
3,3-(Ethylenedioxy)eudesm-ll(12)-eno-13,6a-lactone (6). A solution of 5 (592 mg, 1.32 mmol) in T H F (25 mL) containing acetic acid (0.200 mL, 3.74 mmol) was treated a t 0 "C with 30% H202(0.90 mL, 8.83 mmol) for 1.5 h. The reaction mixture was poured into a cold saturated aqueous solution of NaHC03 (40 mL) and extracted with ethyl acetate (5 X 25 mL). The combined extracts were washed with a saturated aqueous solution of NaCl (3 X 50 mL), dried (Na2S04),and concentrated to give 382 mg of crude crystalline product, which was recrystallized from ethyl acetate to give spectroscopically pure 6 as colorless crystals (359 mg, 93%). The analytical sample of 6 was obtained by the recrystallization from a mixture of benzene, ether, and hexane (1:l:l) as prisms: mp 161-163 "C; IR (KBr) 1775 cm-'; 'H NMR (90 MHz) 6 0.98 (3 H, 9, Clo-CH,), 1.08 (3 H, d, J = 6.0 Hz, C,-CH3), 1.73 (1H, dd, J = 10.5, 10.5 Hz, C5-H), 2.02 (1H, dq, J = 10.5, 6.0 Hz, C4-H), 2.49 (1H, ddddd, J = 10.5, 10.5, 3.0, 3.0, 3.0 Hz, C,-H), 3.78 (1H, dd, J = 10.5, 10.5 Hz, C6-H), 3.96 (4 H, br s, OCH2CH20), 5.34 (1 H, d, J = 3.0 Hz, C12-Ha), 6.01 (1 H, d, J = 3.0 Hz, C12-Hb); 13C NMR 6 12.01 (q, C15), 18.1 (4, C14), 21.5 (t, CB), 31.0 (t, Cz), 36.6 (S, Clo), 39.0 (t,C9), 39.2 (d, C4), 40.2 (t, Cl), 50.8 (d, C5 or Cy),51.6 (d, C5or C,), 65.1 (t,OCH2CH20),83.4 (d, ce),110.4 (5, c3),116.4 (t, c12),139.3 (9, Cii), 170.6 (9, c13); MS, m/e (relative intensity) 292 (M', loo), 99 (45); [U]"OD+29.6" (c 2.93, CHCl,). Anal. Calcd for C17H2404: C, 69.83; H, 8.27. Found C, 69.79; H, 8.41. (11S)-3-Oxoeudesm-ll(12)-eno-13,6a-lactone (7). A mixture of 6 (52 mg, 0.18 mmol), ethanol (4.5 mL), and a 20% aqueous solution of acetic acid (4.5 mL) was refluxed (bath temperature 85 "C) for 4 h. The mixture was cooled, poured into a saturated aqueous solution of NaCl(20 mL),and extracted with ethyl acetate (4 x 10 mL). The combined extracts were washed successively with a saturated aqueous solution of NaHC03 (2 X 10 mL) and a saturated aqueous solution of NaCl(3 X 10 mL), dried (Na$04), and concentrated to give spectroscopically pure 7 (44 mg, 100%) as a colorless crystalline material (mp 153-156 "C), which was recrystallized from ethyl acetate to give colorless needles: mp 171-173 "C; IR (KBr) 1762,1704 cm-'; 'H NMR (90 MHz) 6 1.18 (3 H, S, CIO-CH3), 1.28 (3 H, d, J = 6.8 Hz, Cd-CHJ, 1.69 (1 H, dd, J = 10.5, 10.5 Hz, C,-H), 2.53 (1H, dq, J = 10.5,6.8 Hz, C,-H),
J. Org. Chem., Vol. 52, No. 21, 1987 4795
Notes Table IV. Pot Test Procedures
disease (pathogen) host rice blast (P. oryzae)
test compounds application method preventive foliar spray a few hours preinoculn
dosage: ppm 500
sheath blight (R. solani)
rice
preventive foliar spray a few hours preinoculn
500
powdery mildew (E. graminis)
wheat
preventive foliar spray a few hours preinoculn
500
damping off (P. aphanidermatum)
cucumber solid drench immed postsowing
dawny mildew (P. grape viticora)
500 20 mL/ Pot
preventive foliar spray 1 day preinoculn
200, 50
curative foliar spray 16-18 h postinoculn
500
late blight (P. infestano)
tomato
preventive foliar spray a few hours preinoculn
500
scab (V. inaequalis)
apple
preventive foliar spray a few hours preinoculn
500
incubation method for disease assessment method of inoculation method development disease severity foliar spray of spore 4 days, high evaluation by suspension humidity, darkness, infection indices 28 "C (0-5) depending on number of brown spots infestation of water 4 days, high evaluation by infection indices with mycelium humidity, darkness, grown in the chaffs 28 "C (0-5) depending on medium size of necrotic lesions dusting of spores 10 days, fluorescent evaluation by lamps, 22 "C infection indices (0-5) depending on rate of leaf coverage of powdery mildew infestation of soil 14 days, greenhouse, evaluation by -25 OC with mycelium infection indices grown in the bran's (0-5) depending on medium number bf infected seedlings foliar spray of spore 3 days, high evaluation by suspension humidity, darkness; infection indices 7 days, under (0-5) depending on natural condition, rate of necrosis greenhouse foliar spray of spore 3 days, high evaluation by suspension humidity, darkness; infection indices 7 days, under (0-5) depending on natural condition, rate of necrosis greenhouse foliar spray of spore 7 days, high evaluation by suspension humidity, darkness, infection indices 20 "C (0-5) depending on rate of necrosis foliar spray of spore 5 days, high evaluation by suspension humidity, darkness, infection indices 15 "C; 10 days, (0-5) depending on fluorescent lamps rate of scab lesions
Active ingredient. 3.87 (1 H, dd, J 10.5, 10.5 Hz, Cs-H), 5.40 (1H, d, J = 3.0 Hz, C21H2603Se: C, 62.22; H, 6.46. Found: C, 62.50; H, 6.62. C1z-Ha), 6.05 (1 H, d, J = 3.3 Hz,CiZ-Hb); 13C NMR 6 13.8 (9, C15), Conversion of 10 t o 7. A solution of 10 (2.924 g, 7.21 mmol) 18.3 (q, C14), 21.4 (t, C8), 36.6 (s, Clo), 37.2 (t, Cg), 39.8 (t, C1 or in T H F (45 mL) containing acetic acid (1mL, 17.5 mmol) was treated at 0 "C with 30% HzOz (5.2 mL, 51 mmol) for 1.5 h. The Cz), 40.6 (t, C1 or Cz), 44.6 (d, C4), 50.1 (d, C5 or C7), 54.0 (d, C5 reaction mixture was poured into a cold saturated aqueolls solution or c7),83.2 (d, c6),117.1 (t, clz), 138.6 (9, c11), 170.1 (9, c13),210.9 (s, C3); MS m / e (relative intensity) 248 (M', 43), 137 (36), 124 of NaHC03 (150 mL) and extracted with ethyl acetate (2 X 150 (28), 110 (64), 82 (22), 58 (20), 43 (48), 28 (73), 18 (100); [O ].' D mL, 2 X 100 mL). The combined extracts were washed with a +39.6" (c 2.26, CHC13). Anal. Calcd for C15H2003: C, 72.55; H, 1M solution of KI (100 mL), a 10% aqueous solution of NazSzO3 8.12. Found: C, 72.47; H, 8.14. (1 X 100 mL), and a saturated aqueous solution of NaCl(3 X 150 (1 1S)-11~-(Phenylseleno)-3-oxoeudesmano-l3,6c~-lactone mL), dried (NaZSO4),and concentrated to give a crystalline material, which was recrystallized from ethyl acetate to give 7 (10). A mixture of 5 (3.375 g, 7.51 mmol) and a 50% aqueous solution of acetic acid (60 mL) was refluxed (bath temperature as colorless prisms (1.672 g, 93%). 120 "C) for 20 min. The mixture was cooled, poured into a 2a-Bromo-3-oxoeudesm-ll( 12)-eno-13,6a-lactone (8). The saturated aqueous solution of NaCl(l50 mL), and extracted with solution of phenyltrimethylammonium perbromide (PTAB;2.167 ethyl acetate (4 X 150 mL). The combined extracts were washed g, 5.76 mmol) in T H F (30 mL) was added dropwise to the stirred solution of 7 (1.364 g, 5.49 mmol) in T H F (50 mL) a t -6 "C over successively with a saturated aqueous solution of NaHC03 (3 X a period of 5 min. The mixture was stirred a t -6 "C for 1h and 300 mL) and a saturated aqueous solution of NaCl(2 X 300 mL), dried (NaZS04),and concentrated to give a crystalline material, then poured into a mixture of a saturated solution of NaHC03 which was recrystallized from ethyl acetate to give pale yellow (100 mL) and 0.2 M aqueous solution of Na2S203(100 mL). The needles: 2.955 g, 97%; mp 170-171 "C; IR (KBr) 2935,1765,1710 mixture was extracted with ethyl acetate (5 X 90 mL). The cm-'; 'H NMR (90 MHz) 6 1.19 (3 H, s, C10-CH3),1.24 (3 H, d, combined extracts were washed with a saturated aqueous solution J = 6.9 Hz, CI-CH,), 1.51 (3 H, 9, Cli-CHJ, 1.65 (1H, t, J = 10.5 of NaCl (3 X 200 mL), dried, and concentrated to give a crude Hz, C5-H), 2.50 (1 H, dq, J = 10.5, 6.9 Hz, C4-H), 4.32 (1H, dd, product as pale yellow crystals, which w, 3 recrystaked from ethyl acetate to give colorless prisms: 1.542 g, 86%; mp 147-8 "C dec; J = 10.5,9.8 Hz, Ce-H), 7.20-7.65 (5 H, m, CsH5); 13CNMR 6 13.8 IR (KBr) 1760,1720,1670,1630 cm-'; 'H NMR (90 MHz) 6 1.28 (9, ci5), 18.2 (9, c14), 20.8 (t,c&, 22.2 (9, Ciz), 36.4 (8, cia), 37.1 (3 H, 9, ClO-CHS), 1.38 (3 H, d, J = 6.5 Hz, C4-CH3), 1.77 (1H, (t, Cg), 39.9 (t, C1 or C2), 40.5 (t, C1 or Cz), 44.8 (d, C4), 48.4 (s, dd, J = 11.3, 10.5 Hz, C5-H), 2.09 (1 H, dd, J = 12.8, 12.8 Hz, CiJ, 53.5 (d, C5), 57.0 (d, C,), 80.9 (d, CB), 124.2 ( 8 , CsH5), 128.9 (d, CsH5), 129.6 (d, C&5), 138.0 (d, CsH5), 176.1 (S, C13), 210.9 Cl-H,), 2.43 (1H, dd, J = 12.8,6.5 Hz, Cl-H,), 2.73 (1 H, dq, J = 11.3, 6.6 Hz, C4-H), 3.87 (1 H, dd, J = 10.5, 10.5 Hz, Cs-H), 4.83 ( 8 , C3); [ a I m D +43.7" (c 1.25, CHCl,); MS m / e (relative intensity) 408 [M+ (%e), 201, 406 [M+ (Y3e), 831, 404 [M+ (7sSe),531, 403 (1H, ddd, J = 12.8, 6.5, 1.2 Hz, C,-H), 5.42 (1H, d, J = 3.0 Hz, [M+ (77Se),181,402 [M+ (76Se),161, 400 [M' (74Se),31, 249 (1001, C1z-Ha),6.08 (1H, d, J = 3.3 Hz, Clz-Hb);13CNMR 6 14.1 (q,C15), 248 (68), 231 (30), 175 (27), 158 (27), 111 (53). Anal. Calcd for 18.8 (9, C14), 21.1 (t, Cs), 39.2 (9, Cia), 39.2 (t, Cg), 44.8 (d, C4),
J. Org. C h e m . 1987,52, 4796-4798
4796
50.1 (d, C5 or C7), 52.8 (d, C5 or C7),53.2 (t, Cl), 54.4 (d, Cz), 82.3 (d, Ce), 117.6 (t, Clz), 138.2 (9, C11), 169.9 (9, C ~ Z 201.2 ), (9, C3); MS (13.5 eV), m l e (relative intensity) 328 [M' (81Br),121 327 [M' (*'Br) - 1,431, 326 [M' (79Br),161, 325 [M' (79Br)- 1,331, 248 (24), 246 (54), 231 (57), 229 (loo), 220 (39), 219 (36); (CY]"D +26.4" (c 1.46, CHC13). Anal. Calcd for C15H1903Br:C, 55.06; H, 5.81. Found: C, 54.82; H, 5.95. Tuberiferin (9). A mixture of 8 (450 mg, 1.38 mmol), Li2C03 (270 mg, 3.65 mmol), and LiBr (205 mg, 2.36 mmol) in anhydrous DMF (15 mL) was stirred a t 123-131 "C for 1.5 h, cooled to room temperature, poured into a saturated aqueous solution of NH4Cl (15 mL), and extracted with ethyl acetate (5 X 20 mL). The combined extracts were washed with a saturated aqueous solution of NaCl(3 X 50 mL), dried (NazSO,), and concentrated to give a crystalline material, which was recrystallized from ethyl acetate to give spectroscopically pure 9 (289 mg, 85%). The analytical sample of 9 was obtained by further recrystallization from ethyl acetate as prisms: mp 179-181 OC;lZIR (KBr)123085,3025,1763, 1665, 1625, 1410, 1250, 1235, 1142,997, 965, 945,830 cm-'; 'H NMR (90 MHz) 6 1.17 (3 H, 9, Cio-CH3), 1.39 (3 H, d, J = 6.8 Hz, C4-CH3), 2.08 (1H, dd, J = 10.5, 12.0 Hz, C5-H), 2.58 (1 H, dq, J = 12.0, 6.8 Hz, C4-H), 3.96 (1H, dd, J = 10.5, 10.5 Hz, C6-H), 5.41 (1H, d, J = 3.0 Hz, C12-Ha),5.85 (1H, d, J = 10.0 Hz, CZ-H), 6.05 (1H, d, J = 3.3 Hz, CiZ-Hb), 6.68 (1 H, d, J = 10.0 Hz, C1-H); 13CNMR 6 14.6 (9,C15), 19.2 (q, C14), 21.1 (t, Cg), 37.1 (t,Cg), 38.4 (s, Clo),41.9 (d, C4),50.1 (d, C5 or C7),52.2 (d, C5 or C7),81.9 (d, Ce), 117.2 (t, C ~ Z 126.4 ), (d, C2), 138.3 (9, CiJ, 158.0 (d, CJ, 170.0 (s, C13), 200.2 (s, CJ; MS (13.5 eV), m l e (relative intensity) 246 (M', loo), 218 (74), 192 (27), 190 (20); [ C X ] ~ ~+12.2" D ( C 2.12, CHC13). Anal. Calcd for C15H1803: C, 73.15; H, 7.37. Found C, 72.98; H, 7.33. When the crude sample of 9 was chromatographed over silica gel and eluted successively with chloroform and ethyl acetate, the eluent with ethyl acetate gave a crystalline material, mp 155-157 0C.13,14 Although the 'H NMR spectrum in CDCl, and IR spectrum in chloroform were identical with those of prisms, mp 179-181 "C, above mentioned, the IR spectrum recorded as KBr disk was apparently different: IR (KBr)133100, 3040, 2950, 1765, 1680,1622,1412,1255,1240,1150,1140,1000,990,950,838 cm-'; IR (CHC13) 3025,2945,1765,1670,1410,1245,1137,998,967,823 cm-l. Synthesis of Tuberiferin (9) from 7 without Purification of 8. The solution of PTAB (255 mg, 0.677 mmol) in T H F (3.0 mL) was added to the stirred solution of 7 (150 mg, 0.605 mmol) in T H F (6 mL) a t -2 "C over a period of 5 min. The mixture was stirred for 1h at this temperature and worked up as usual to give 216 mg of crude 8. The mixture of this crude 8 (216 mg), Li2C03 (131 mg, 1.77 mmol), and LiBr (98 mg, 1.13 mmol) in anhydrous DMF (6.5 mL) was stirred at 123-131 "C for 1.5 h and worked up as usual to give a crystalline material (183 mg), which was purified by column chromatography [Merck, silica gel, 70-250 mesh, 15 g, AcOEtCHC1, (1:9)]to give spectroscopicallypure 9 (130 mg, 87% overall yield from 7). Cell Growth Inhibitory Activity of Compounds to Murine Lymphocytic Cell (P388) in Vitro. Murine lymphocytic leukemia cells (P388) were incubated with compounds a t 37 "C in humidified atmosphere of 5% COz for 48 h. After incubation, the cell number was counted with a Coulter counter (Model ZBI, Coulter Electronics, Inc., Haialeah, FL), and the cell growth inhibition ratio (%) was calculated according to cell growth inhibn ratio (%) =
( -1
::::)x100
where T = cell count after culture with compound, C = cell count after culture without compound, and Co = cell count at the start of culture. (12) Melting point and IR (KBr) data recorded here are in good accordance with those reported by K. Yamakawa et (13) Melting point and IR (KBr) data recorded here are in good accordance with those reported by J. B. Barrera et al.9 (14) A possible explanation is that (+)-tuberiferin has two crystal forms whose melting points and IR spectra in KBr disk are different from each other. 0022-326318711952-4796$01.50/0
Plant Growth Regulation Activity of Compounds. The compounds (15 mg)dissolved in 0.3 mL of solvent (acetoneTween 80, 1O:l) were diluted with water to give test solutions. Three kinds of seeds, E. frumentacea, B. juncea, and C. sativus, were sown in a Petri dish (10 cm X 15 cm) containing 10 mL of the test solution and incubated under light (4000 lx) at 27 "C for 10 days. The germination of seeds and the growth of seedlings were observed and examined. Effects of compounds on plants were expressed as four rating scales (+++, serious or complete inhibition; ++, obvious effect, +, slight detectable effect; -, no effect). Preventive or Curative Activity of Compounds in Controlling Crop Diseases. The diseases and the test methods are shown in Table IV. Test samples, which were formulated as emulsifiable in water, were applied by spraying to the plants or drenching to soil before or after inoculation. The plants were inoculated with spores or hypha of fungal pathogens. After incubation, disease severity of test plants was observed under desirable conditions for 4-15 days. The activity was expressed as a score from 0 through 5 (0, 0-29%; 1, 30-49%; 2, 50-69%; 3, 7 0 4 9 % ; 4, 90-99%; 5, 100%; controlled vs. untreated).
Acknowledgment. We express our thanks to Professor S. Yamaguchi for a loan of a polarimeter and for his help in the measurement of optical rotation. Registry No. 1, 481-06-1; 2, 14804-46-7; 3, 13902-54-0; 4, 66767-50-8; 5, 66726-14-5; 6, 110193-06-1; 7, 66726-13-4; 8, 92214-65-8; 9, 18375-00-3; 10, 99297-23-1; 11, 62326-48-1; 12, 39813-41-7.
Activation of Zinc by Trimethylchlorosilane: An Improved Procedure for the Preparation of /?-Hydroxy Esters from Ethyl Bromoacetate and Aldehydes or Ketones (Reformatsky Reaction) GBrard Picotin and Philippe Miginiac* Laboratoire de Chimie des Organom&talliques,U A 574 CNRS, Universite de Poitiers, 86022 Poitiers Cedex, France Received March 11, 1987
The Reformatsky reaction is the Reformatsky generally applicable procedure for converting aldehydes and ketones to @-hydroxyesters (Scheme I).1-5
Scheme I RCOR'
e
+ BrCH2COOC2H,
(1) Zn
RC(OH)(R')CH2COOC2H, Normally the reaction is carried out at reflux temperatures by adding carbonyl substrate and bromo ester simultaneously to a suspension of zinc in an appropriate solvent, generally benzene or an ether-benzene mixture. Numerous variations have been proposed in order t o improve both the yield and the purity of the hydroxy esters formed.6-11 (1)Shriner, R. L. Org. React. (N.Y.) 1942, 1 , 1. (2) Rathke, M. W. Org. React. (N.Y.) 1976,22, 423. (3) Gaudemar, M. Organomet. Chem. Rev. Sect. A 1972,8, 183.
(4) Henecka, H. In Methoden der Organischen Chemie, 4th ed.; Muller, E., Ed.; Georg Thieme Verlag: Stuttgart, 1952;Vol. VIII, p 511. (5) Nutzel, K. In Methoden der Organischen Chemie, 4th ed.; Miiller, E., Ed.; Georg Thieme Verlag: Stuttgart, 1973, Vol. XIII/2a, p 809. (6) Use of a trimethyl borate-tetrahydrofuran mixture as solvent: Rathke, M. W. J. Org. Chem. 1970,35, 3966. (7) Use of a zinc-copper couple: Santaniello, E.; Manzocchi, A. Synthew 1977, 698.
0 1987 American Chemical Society
